摘要
以实验室前期筛选得到的分枝杆菌(Mycobacterium sp.LY-1)为出发菌株,采用等离子诱变(ARTP)技术选育出能高效转化植物甾醇为重要甾体药物中间体9α-羟基雄甾-4-烯-3,17-二酮(9α-OH-AD)的菌株,并对其转化工艺进行优化。经过ARTP诱变选育得到遗传稳定性较好的分枝杆菌突变株Mycobacterium sp.C33,当底物植物甾醇投料浓度为15 g/L时,转化生成9α-OH-AD的摩尔得率达到15.5%,较原始菌株提高34.8%。继而采用正交实验设计方法,对突变菌的发酵培养基组分进行优化,并建立了油水双相转化体系,进一步提高了突变菌株C33的产物摩尔得率,最高达到47.0%,较优化前(15.5%)提高了2倍。
In order to improve transformation efficiency of phytosterols into 9α-hydroxylation of 4-androstene-3,17-dione(9α-OH-AD) by Mycobacterium sp. LY-1, we studied the strains breeding using atmospheric and room temperature plasma(ARTP) technology and optimized their conversion process. A high production strain named C33 with a good genetic stability was selected and the product molar yield reached to 15.5%, 34.8% higher than that of original strain with 15 g/L phytosterols. Furthermore, the fermentation medium was optimized through the design of orthogonal experiment. Besides, oil-water bidirectional transformation system was set up to improve the 9α-OH-AD molar yield of mutant strain C33. With adding 12 mL soybean oil to each 1 g phytosterols, the molar yield of 9α-OH-AD reached 47.0%, which increased twice than that of control(15.5%).
出处
《生物工程学报》
CAS
CSCD
北大核心
2017年第7期1198-1206,共9页
Chinese Journal of Biotechnology
基金
国家高技术研究发展计划(863计划)(No.2011AA02A211)
江苏省普通高校学术学位研究生科研创新计划项目(No.KYLX16_0826)资助~~
关键词
分枝杆菌
植物甾醇
9α-OH-AD
等离子诱变
生物转化
Mycobacterium sp
phytosterol
9α-OH-AD
atmospheric and room temperature plasma
biotransformation
作者简介
Corresponding author： Hui Li. Tel： ＋86-510-85326883; Fax： ＋86-510-85328177; E-mail： lihui@jiangnan.edu.cn
